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1. 发明申请

WO2015165879A1 DEVICE AND METHOD FOR LARGE VOLUME TRANSFECTION 审中-公开
标题翻译：用于大容量转移的装置和方法
公开(公告)号：WO2015165879A1
公开(公告)日：2015-11-05
申请号：PCT/EP2015/059150
申请日：2015-04-28
申请人： LONZA COLOGNE GMBH
发明人： ALTROGGE, Ludger , GLEISSNER, Timo , HEINZE, Andreas , HERMSMEIER, Sven
IPC分类号： C12M1/42 , A61N1/30 , C12N15/87 , G01N35/10
CPC分类号： C12N13/00 , A61N1/0412 , C12M35/02 , C12N15/87
摘要： The invention relates to a device (1) for applying an electric field to a suspension of cells, cell derivatives, organelles, sub-cellular particles and/or vesicles, comprising at least one chamber (6) which comprises at least two electrodes (4), and at least one separating element (13) which is movable within the chamber (6) between two terminal points (14, 15) and, if it is in a position between the terminal points (14, 15), separates at least one first compartment (26) of the chamber (6) from at least one second compartment (27) of the chamber (6). According to the invention each compartment (26, 27) is designed to hold the suspension and comprises at least one port (7, 8, 10, 11) for charging or discharging the suspension, so as to discharge an aliquot of the suspension from the chamber (6) and at the same time charge a further aliquot of the suspension into the chamber (6), wherein the separating element (13) is moved in a second direction opposite to a first direction, and wherein the separating element (13) separates the aliquots from each other.
摘要翻译：本发明涉及一种用于向细胞，细胞衍生物，细胞器，亚细胞颗粒和/或囊泡的悬浮液施加电场的装置（1），包括至少一个包含至少两个电极（4）的腔室（6））和至少一个分离元件（13），其可在所述腔室（6）内在两个端点（14,15）之间移动，并且如果其处于所述端点（14,15）之间的位置，则至少分离来自腔室（6）的至少一个第二隔室（27）的腔室（6）的第一隔室（26）。根据本发明，每个隔室（26,27）被设计成保持悬架并且包括至少一个端口（7,8,10,11），用于对悬架进行充电或排出，以将悬架的等分试样从同时将悬浮液的另一等分装入室（6），其中分离元件（13）在与第一方向相反的第二方向上移动，并且其中分离元件（13）将等分试样彼此分离。

2. 发明申请

WO2015165881A1 DEVICE AND METHOD FOR LARGE VOLUME TRANSFECTION 审中-公开
标题翻译：用于大容量转移的装置和方法
公开(公告)号：WO2015165881A1
公开(公告)日：2015-11-05
申请号：PCT/EP2015/059152
申请日：2015-04-28
申请人： LONZA COLOGNE GMBH
发明人： ALTROGGE, Ludger , GLEISSNER, Timo , HEINZE, Andreas , HERMSMEIER, Sven
IPC分类号： C12M1/42 , A61N1/30 , C12N15/87 , G01N35/10
CPC分类号： C12N13/00 , A61N1/0412 , C12M35/02 , C12N15/87
摘要： The invention relates to a device for applying an electric field to a suspension of cells, comprising at least one chamber which comprises at least one internal space (40) for holding the suspension, the internal space (40) comprising at least two segments (41, 42), wherein each segment (41, 42) comprises at least one electrode (43, 44) and wherein neighboring electrodes (43, 44) are separated from each other by at least one gap (47) which is at least partially filled with an insulating material (46), and wherein the edges of the electrodes (43, 44) facing each other within the internal space (40) are rounded. Rounding the electrodes' edges facing a neighboring electrode results in a significant reduction of field gradients and thus even of the risk of arcing. The invention further concerns a method,wherein voltage is applied to at least one active electrode (43, 44) while the electrodes (43, 44, 45) or electrode segments next and/or opposite to the active electrode (43, 44) are set to ground potential. Setting neighboring electrodes that surround the active electrode to ground potential results in decreased scattering of the electric field within the internal space so that the electrically active area is locally limited and the field lines are focused near the active electrode and thus control of the process is enhanced.
摘要翻译：本发明涉及一种用于向电池悬浮液施加电场的装置，包括至少一个腔室，该腔室包括用于保持悬架的至少一个内部空间（40），所述内部空间（40）包括至少两个段（41 ，42），其中每个段（41,42）包括至少一个电极（43,44），并且其中相邻电极（43,44）通过至少一个间隙（47）彼此分开，所述间隙至少部分地被填充具有绝缘材料（46），并且其中在内部空间（40）内彼此面对的电极（43,44）的边缘是圆形的。将电极的边缘朝向相邻电极四舍五入会导致现场梯度的显着降低，从而导致电弧的风险。本发明还涉及一种方法，其中电压被施加到至少一个有效电极（43,44），而与有源电极（43,44）相邻和/或相对的电极（43,44,45）或电极段是设定为地电位。将围绕有源电极的相邻电极设置为接地电位导致电场在内部空间内的散射减少，使得电活性区域局部受限并且场线聚焦在有源电极附近，从而增强了该过程的控制。

3. 发明授权

EP1427837B1 MODULARE TRANSFEKTIONSSYSTEME AUF DER BASIS VON NUKLEOPROTEINFILAMENTEN 有权
标题翻译：模块化转基于NUKLEOPROTEINFILAMENTEN
公开(公告)号：EP1427837B1
公开(公告)日：2011-10-26
申请号：EP02704571.5
申请日：2002-01-10
申请人： Lonza Cologne GmbH
发明人： SCHMIDT, Hanns-Martin , ALTROGGE, Ludger , LENZ, Dietmar , RIEMEN, Gudula , BROSTERHUS, Helmut , LORBACH, Elke , HELFRICH, Juliana , HEIN, Katharina , GREMSE, Marion , MALES, Tatjana , CHRISTINE, Rainer , SIEBENKOTTEN, Gregor , ORTMANN, Bodo , TURBANSKI, Tamara , KLAES, Andrea
IPC分类号： C12N15/87 , C07K14/47 , C07K19/00 , A61K48/00 , G01N33/50 , C12Q1/68
CPC分类号： C07K14/47 , A61K48/00 , C07K2319/035 , C07K2319/09 , C07K2319/10 , C07K2319/21 , C12N15/87
摘要： The invention relates to a method for the transfection of cells using at least one protein capable of forming nucleoprotein filaments. Said protein is first modified by at least one functional constituent which influences at least one step of the transfection, and the nucleic acid to be transfected is then loaded with the modified protein. The nucleic acid and the protein form a filamentary complex, and said complex is then added to the cells to be transfected. The invention also relates to a transfection agent of nucleoprotein filaments (NPF), at least one protein forming a nucleoprotein filament being modified by at least one functional constituent for the transfection. The invention further relates to the use of the inventive transfection agent for producing a medicament for the gene therapeutic treatment of humans and animals. Finally, the invention relates to corresponding pharmaceutical preparations, especially for use in gene therapy, and the use of such transfection agents in the form of constituents in kits. Fig. 20. 1. DNA 2. protein building blocks 3. charged DNA 4. functions 5. association with the cell surface 6. passage through the membrane 7. emergence from the endosomes 8. transport into the nucleus 9. other function 10. cell membrane 11. nucleus membrane 12. separation in nucleus 13. expression

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